KR20160076821A - Fermented vegetable oils having high free essential unsaturated fatty acids and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a fermented vegetable oil and a manufacturing method for the same for the prevention of skin itchiness, atopic dermatitis, and cell membrane damage and for moisturization based on the production of fermented vegetable oil rich in free essential unsaturated fatty acid based on vegetable oil fermentation with rhizopus microorganism culture concentrate. The present invention is obtained by fermenting the vegetable oil selected from the group consisting of borage oil, moringa oil, evening promise seed oil, camellia oil, olive oil, sunflower seed oil, and a mixture of at least two thereof with the rhizopus microorganism culture concentrate. Also provided is a moisturizing skin external preparation or cosmetic material composition containing the fermented vegetable oil and preventing skin itchiness, atopic dermatitis, and skin cell membrane damage.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a fermented vegetable oil having a high content of free essential unsaturated fatty acids and a method for producing the fermented vegetable oil,

The present invention relates to a fermented vegetable oil having a high content of free essential unsaturated fatty acids and a method for producing the fermented vegetable oil. More specifically, the present invention relates to a fermented vegetable oil having a high content of free essential unsaturated fatty acids and fermenting the vegetable oil with a culture concentrate of Rizophus genus microorganism To a plant fermented oil for improving skin itching and atopic dermatitis, moisturizing effect, and prevention of cell membrane damage, and a method for producing the same.

Recently, facial oil has received much attention. This is because facial oils restore the oil and water film of the skin and enhance the moisturizing power.

The effect of using cosmetic oil is to form a hydrophobic coating on the skin surface to prevent penetration of harmful substances from the outside and to suppress the evaporation of moisture on the skin surface. It is also used as a solvent for chemical sunscreen agents and fat-soluble vitamins. Fat can be a precursor to ceramides and vitamins, depending on the type of fatty acids they contain. Thus, interest in essential unsaturated fatty acids in oils is also increasing.

Polyunsaturated fatty acids, which are essential for normal development and maintenance of human homeostasis in humans, but which can not be synthesized in the body, are called essential unsaturated fatty acids. In general, Gamma-Linolenic Acid (GLA) is made in the body from linoleic acid (LA), another essential unsaturated fatty acid. Gamma linolenic acid is an omega-6 fatty acid that inhibits inflammation and allergic reactions in the body, regulates the immune response, and plays an important role in cell membrane formation and maintenance. Oleic acid (OA) lowers the level of LDL-cholesterol, which is a harmful low-density cholesterol, while it increases HDL-cholesterol, a good high-density cholesterol in the body, to prevent hypertension and heart disease. It is also known to be effective in preventing cancer and declining memory due to aging.

The top layer of skin is composed of fatty acids, cholesterol and ceramide. The reason why essential unsaturated fatty acid is important is because it consists of ceramide or sphingosine which plays a role of skin moisturizing factor and lipid structure which is combined with essential unsaturated fatty acid. Fatty acids are a key component of skin health. Unless the linoleic acid (LA), an essential unsaturated fatty acid forming the ceramide layer and epidermal membrane of the skin, is normally converted to gamma linolenic acid, it causes excessive moisture loss through the epidermis, resulting in failure to maintain healthy keratinocytes and dry skin.

In addition, the skin membranes of the dermal layer are composed of essential unsaturated fatty acids. If the elastic fatty acid is insufficient, skin dryness, nail cracking, skin dandruff, seborrheic dermatitis and hair loss occur. The skin cells produced from the dermis are subjected to a keratinization process in which they disappear into keratinocytes at a period of about 4 weeks. When the essential unsaturated fatty acids are insufficient, the production of dermal cells becomes difficult, causing skin troubles and inflammation do.

However, since the fat in the form of glycerol and fatty acid bonds is difficult to absorb quickly into the skin, it is necessary to have a form of free unsaturated fatty acid that is easily absorbed. To produce the essential unsaturated fatty acids liberated, a process such as fermentation is required. Conventional methods of fermenting oil include a method of germinating oil seeds, inoculating the microorganism by solid fermentation, separating and extracting lipids, and adding microorganisms to vegetable oil for fermentation. However, it takes a long time to perform solid fermentation, and since the production efficiency of the free essential unsaturated fatty acid is lowered and the process of extracting the lipid is required, there is a disadvantage that the process ratio is increased, and the method of directly adding the microorganism to the vegetable oil, , And it has a disadvantage that it is not suitable for use in cosmetics. In addition, a common disadvantage of solid fermentation and direct fermentation of microorganisms is that it is difficult to completely remove the microorganisms used in the fermentation, so there is a possibility of re-contamination.

On the other hand, lipase is an enzyme that hydrolyzes lipid to fatty acid (Fatty acid) and glycerol (Glycerol). Lipase is somewhat different in specificity as it is derived from microorganisms and from the pancreas of animals such as pigs. The characteristics of some bacteria that produce lipase are as follows. Candida antarctica is an industrially important resource for the production of lipases. In addition, Rhizopus oryzae , including Rhizopus oryzae , is harmless to humans and is widely used in the food industry and food, and is known to produce a large amount of lipase.

Accordingly, the present inventors have found that a fermentation period is shortened by supplementing the disadvantages of solid fermentation and microbial fermentation, there is no odor that is unsuitable for use in cosmetics, no possibility of re-contamination, and a large amount of lipase is produced to efficiently produce essential unsaturated fatty acid It is possible to increase the content of free essential unsaturated fatty acids by fermenting a large amount of vegetable oil by using a culture concentrate of a microorganism known as a microorganism, especially a culture concentrate of a microorganism of the genus Rizophus, to regenerate a damaged skin cell membrane, to exert an excellent moisturizing effect, The present invention has been completed by producing an oil having an atopic dermatitis-improving effect.

It is an object of the present invention to provide a fermented vegetable oil in which a culture concentrate of microorganisms of the genus Rizophtus is added to a vegetable oil and the oil is fermented to contain a large amount of free essential unsaturated fatty acid.

Another object of the present invention is to provide a method for producing a fermented vegetable oil in which a culture concentrate of microorganisms of the genus Rizophus is added to a vegetable oil and the oil is fermented to contain a large amount of free essential unsaturated fatty acid.

Another object of the present invention is to provide a pharmaceutical composition for external skin application for preventing skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement containing the above fermented vegetable oil as an active ingredient .

It is still another object of the present invention to provide a cosmetic composition for prevention of skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement containing the fermented vegetable oil as an effective ingredient.

It is still another object of the present invention to provide a quasi-drug for preventing skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement containing the fermented vegetable oil as an effective ingredient.

The fermented vegetable oil according to the present invention is obtained by cultivating a plant oil selected from the group consisting of Borage Oil, Moringa Oil, Evening Primrose Oil, Camellia Oil, Olive Oil, Sunflower Seed Oil and a mixture of two or more thereof, Obtained by fermentation with a concentrate.

The culture concentrate of the Rizophus genus microorganism is inoculated with the microorganism of the genus Rizophus in an amount of 10 vol.% Relative to the medium volume and cultured under culture conditions at pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, It may be a precipitate obtained by salting out the supernatant obtained by centrifuging at 1 to 10 DEG C at 800 to 1200 rpm.

The medium may be PDB (Potato Dextrose Broth) containing 2 to 10% by weight of potato starch, 10 to 20% by weight of dextrose and water as a remainder.

The PDB may further comprise 0.1 to 10% by weight of oil based on the total weight of the PDB.

The culture time under the above culture conditions may be within a range of 80 to 150 hours.

Wherein the fermentation of the vegetable oil is carried out by adding a culture concentrate of the microorganism of the genus Rizopus as a remainder to 10 to 80% by weight of preheated vegetable oil at a temperature within a range of 10 to 40 DEG C, Fermentation under fermentation conditions including stirring at a temperature within a range of 25 to 50 DEG C and within a range of 50 to 500 rpm. As described above, the fermentation-completed oil and the culture concentrate can be separated and separated by centrifugation, and the centrifugation and filtration processes can be easily understood by those skilled in the art. You can.

The microorganism of the genus Rizophus may preferably be Rhizopus oryzae (microorganism resource center accession number KCTC6106).

The present invention also provides a method for producing a fermented vegetable oil by fermenting a vegetable oil, comprising the steps of: (1) mixing 10 to 80 weight of a vegetable oil preheated to a temperature within a range of 10 to 40 ° C % Of a culture concentrate of the microorganism of the genus Rizopus as a remaining amount; And (2) fermentation under fermentation conditions including a pH within a range of 5.0 to 8.0, a temperature within a range of 25 to 50 DEG C, and a stirring within a range of 50 to 500 rpm, Wherein the oil is selected from the group consisting of borage oil, moringa oil, evening primrose oil, camellia oil, camellia oil, olive oil, sunflower seed oil and mixtures of two or more thereof.

The culture concentrate of the Rizophus genus microorganism is obtained by inoculating the microorganism of the genus Rizophus into the medium in an amount of 10% by volume based on the medium volume and culturing under culture conditions of pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, Followed by salting out the supernatant obtained by centrifugation at 800 to 1200 rpm at 1 to 10 < 0 > C.

The medium may be a Potato Dextrose Broth (PDB) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose, and water as a remainder.

The culture time under the above culture conditions may be within a range of 80 to 150 hours.

The microorganism of the genus Rizophus may preferably be a Rythopus oryzae.

According to the present invention, there is provided a pharmaceutical composition for external skin for preventing skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement containing the above fermented vegetable oil as an effective ingredient.

According to the present invention, there is provided a cosmetic composition for preventing skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement containing the fermented vegetable oil as an effective ingredient.

According to the present invention, there is also provided a quasi-drug for preventing skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement containing the fermented vegetable oil as an effective ingredient.

According to the present invention, a fermented vegetable oil rich in free essential unsaturated fatty acids can be produced by fermenting vegetable oil with a culture concentrate of Rizophus genus microorganism to produce a vegetable fermentation oil for improving skin itching and atopic dermatitis, moisturizing effect, There is an effect that the manufacturing method is provided.

1 is a graph showing the NO production inhibitory effect of a fermented vegetable oil prepared by fermenting Borage oil, Moringa oil, Evening Primrose oil and Camellia oil according to the present invention in comparison with the control group and Comparative Examples 1 to 4.
FIG. 2 is a graph showing the COX-2 activity inhibitory effect of the fermented vegetable oil prepared by fermenting Borage oil, Moringa oil, Evening Primrose oil and Camellia oil according to the present invention, in comparison with Comparative Examples 1 to 4.
FIG. 3 is a graph showing inhibitory effect on histamine production of fermented vegetable oils prepared by fermenting Borage oil, Moringa oil, Evening Primrose oil and Camellia oil according to the present invention, in comparison with the control group and Comparative Examples 1 to 4.
FIG. 4 is a graph showing the effect of inhibiting the activity and inhibiting degranulation of mast cells in fermented vegetable oils prepared by fermenting Borage oil, Moringa oil, Evening Primrose oil and Camellia oil according to the present invention. 4 is an effect on the activity and degranulation of mast cells.
FIG. 5 is a graph showing inhibition of the activity of mast cells and inhibition of degranulation of fermented vegetable oil prepared by fermenting Borage oil, Moringa oil, Evening Primrose oil and Camellia oil according to the present invention.
FIG. 6 is a graph showing cell stability of fermented vegetable oil prepared by fermenting Borage oil according to the present invention. FIG.
FIG. 7 is a graph showing cell stability of a fermented vegetable oil prepared by fermenting Moringa oil according to the present invention. FIG.
FIG. 8 is a graph showing cell stability of fermented vegetable oil prepared by fermenting evening primrose oil according to the present invention. FIG.
9 is a graph showing the cell safety of the fermented vegetable oil produced by fermenting the camellia oil according to the present invention.

Hereinafter, the present invention will be described in detail.

The fermented vegetable oil according to the present invention is obtained by cultivating a plant oil selected from the group consisting of Borage Oil, Moringa Oil, Evening Primrose Oil, Camellia Oil, Olive Oil, Sunflower Seed Oil and a mixture of two or more thereof, And is obtained by fermentation with a concentrate.

The vegetable oil to be fermented according to the present invention may be selected from the group consisting of borage oil, moringa oil, evening primrose oil, camellia oil, olive oil, sunflower seed oil and mixtures of two or more thereof. It is not intended to be limited thereto, and it should be understood that vegetable oils containing a large amount of unsaturated fatty acids, preferably among vegetable oils, are also within the scope of the present invention.

The Borax oil is known to be rich in gamma linoleic acid and is known to be effective against hypoglycemia, antiinflammation, weight loss, osteoporosis, and has been reported to reduce menopausal syndrome and menstrual syndrome in women.

Moringa oil is known to be one of the three major oils that are good for skin aging, along with argan oil and saccain oil. Especially, it is used for massage because it has excellent skin cleansing function. It keeps moisture of skin well and prevents drying. It is also rich in vitamins and amino acids, making it well suited to rough and aged skin.

The evening primrose oil has been used as indigenous medicines by indians gathering wild evening primrose and grinding it whole. It has been studied to have effects such as lowering blood cholesterol, blood pressure control, and atopic dermatitis relief.

The camellia oil is a fatty oil extracted from camellia Japonica L. of Camellia Japonica L. and the seeds of the same plant. It is used for cream, milk and the like, and is widely used for hair.

The olive oil is oil of 30 to 70% contained in the fruit of an olive tree (olive tree), and is squeezed by a pressing method and an extraction method. The main component of the fatty acid is oleic acid, an unsaturated acid, with a content of about 65 to 85%, and palmitic acid as the main fatty acid. Edible oil is mainly used for salad oil and oil pickling, and is widely used for cooking mayonnaise, salad dressing, frying, and frying. It is used worldwide as a finest product among edible oils, and it is most widely used in cooking, especially in Greece, España and Italy. It is also used in cooking, as well as in make-up and treatment. Olive oil is effective in treating various dermatitis and ulcers and improving skin. In particular, the Mediterranean diet using olive oil is effective in preventing and treating hypertension, diabetes, heart disease, and so on. The essence of the Mediterranean diet is often to eat olive oil, so health practices that once took a spoonful of olive oil every morning in Japan were once popular. The effects of olive oil so far known are:

(1) Prevention of aging: Olive oil contains a large amount of unsaturated fatty acids and antioxidants such as vitamin E, tocopherol, polyphenol, etc., and thus has an excellent effect in preventing aging.

(2) Prevention of Cancer: Antioxidant-rich olive oil is a great help because the cause of cancer is the oxidation of cells. It is especially effective in preventing digestive system cancer.

(3) Prevent cholesterol to prevent adult diseases: rich in antioxidants, such as unsaturated fatty acids and tocopherol, inhibits the formation of harmful cholesterol, prevents arteriosclerosis. It is also effective in preventing heart disease and hypertension.

(4) help digestion and cure stomach cramps: regulate the secretion of gastric juice to treat ulcers, alleviate abdominal pain and help digestion.

(5) Helps liver function: It promotes secretion of beneficial cholesterol (HDL) from the liver, helps liver function and prevents gallstones. HDL can also be a source of energetic agents.

(6) Treating and preventing constipation: The vegetable fat component promotes the bowel movement of the large intestine to treat constipation and soften the stool.

(7) Prevention of diabetes: It is effective to prevent diabetes by controlling blood sugar.

(8) Helps children grow and develops the brain: Vitamins and essential minerals help the bones and brain development of the fetus and growing children.

(9) It is effective for dieting: Olive oil does not contain trans fatty acid which is the cause of obesity, so it is effective for prevention of obesity and diet.

(10) Protecting the skin: Vitamin E and provitamin in olive oil prevent skin aging and release wastes and toxins, making the skin clear and shiny.

(11) Soothe skin troubles: Olive oil has its own sterilizing ability. If you apply lightly to the place where the skin trouble occurs, the trouble calms down.

The sunflower seed oil is an oil obtained from sunflower seeds. The sunflower seed efficacy is an effect to prevent anemia due to folate. Especially, it is a good food for pregnant women who is easily deficient in folic acid. It is a food which maintains skin, antioxidant effect to prevent aging, And phytosterol and magnesium (Sunflower seed contains 18 times more folic acid than phytosterol and tomato), it helps to release bad cholesterol accumulated in blood vessels and it is good to prevent heart disease, It is known that it is an effective food for prevention of hardening, and the abundant magnesium ingredient is known to have an effect of alleviating and preventing symptoms of hypertension by preventing the rise of blood pressure. It is also known to have anti-inflammatory, antioxidant, and cholesterol-removing effects. In addition, the minerals in sunflower seed oil are classified into massive elements and trace elements, among which sunflower seeds contain a large amount of selenium. The average daily selenium requirement of adult men and women is 42 mg per day. The recommended intake is 50 ㎎. Although only a small amount is enough in the human body, the antioxidant effect to prevent the aging of the body is excellent. Especially, It is said that vitamin E is as high as 2000 times. Selenium is known to be good for the prevention of skin aging and the treatment of inflammation, which is also known to be due to its excellent antioxidant activity.

According to the present invention, a culture concentrate of a microorganism of the genus Rizophus is used for fermenting the vegetable oil.

The culture concentrate of the Rizophus genus microorganism is obtained by inoculating the microorganism of the genus Rizophus into the medium in an amount of 10% by volume based on the medium volume and culturing under culture conditions of pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, It may be a precipitate obtained by salting out the supernatant obtained by centrifugation at 800 to 1200 rpm at 1 to 10 DEG C, and by using the culture concentrate of the Ricofus aureus microorganism, the concentration of the unsaturated essential fatty acid It has been confirmed by the present inventors that the production amount can be greatly increased.

At this time, the medium may be potato starch, PDB (Potato Dextrose Broth) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose and water as a remainder. The culture using the PDB may preferably be performed at a pH of 5.1 + - 0.2 and under exhalation conditions, and the PDB may preferably further comprise 0.1 to 10% by weight of oil based on the total weight of PDB .

Particularly, according to the present invention, it was confirmed by the present inventors that the fermentation culture time of the cultured concentrate of the microorganism of the genus Rizophus is the best when the microorganism is cultured within a range of 80 to 150 hours.

The Rhizopus sp. Microorganism is one of the fungi of the fungus Mucorales. It is a branch of the stolon that spreads sideways and extends to the point where it touches the medium. node, and one to several sporangia and rhizoids. Sporangia are large, usually black. The columella is hemispheric, mycelium occurs in large quantities, and it also grows long to fill the Petri dish lid. In essence, two spousal cysts contact and fuse to form junction spores. Soil, fruit, etc., and rarely human, animal, or some species of plants are pathogenic. It has a strong starch sugar strength and a very small amount, but it has alcohol fermentation ability and proteolytic ability and also has a strong ability to produce organic acids such as lactic acid and fumaric acid. For example, it is used for the brewing of Chinese sake and the production of rice wine in Southeast Asia. R. delemar and R. japonicus are important as amylose for the production of alcohol. R. nigricans is also known to occur in fruits such as peaches and strawberries, cereals, and bread, and also causes sweet potato disease, but it is also known to have a strong fumaric acid-producing ability.

The culture concentrate of the microorganism of the genus Rizophus contains a lipase, and the lipase means an enzyme which decomposes into glycerol and fatty acid.

Wherein the fermentation of the vegetable oil is carried out by adding a culture concentrate of the microorganism of the genus Rizopus as a remainder to 10 to 80% by weight of preheated vegetable oil at a temperature within a range of 10 to 40 DEG C, At a temperature within a range of 25 to 50 DEG C and within a range of 50 to 500 rpm, and the microorganism of the genus Rizophus is preferably Rhizopus oryzae .

The present invention also provides a method for producing a fermented vegetable oil by fermenting a vegetable oil, comprising the steps of: (1) mixing 10 to 80 weight of a vegetable oil preheated to a temperature within a range of 10 to 40 ° C % Of a culture concentrate of the microorganism of the genus Rizopus as a remaining amount; And (2) fermentation under fermentation conditions including a pH within a range of 5.0 to 8.0, a temperature within a range of 25 to 50 DEG C, and a stirring within a range of 50 to 500 rpm, Wherein the oil is selected from the group consisting of borage oil, moringa oil, evening primrose oil, camellia oil, olive oil, sunflower seed oil and mixtures of two or more thereof.

The mixing step (1) comprises adding a culture concentrate of the microorganism of the genus Rizopus as a remainder to 10 to 80% by weight of preheated vegetable oil at a temperature within a range of 10 to 40 캜.

The fermentation step of (2) is performed by fermentation under a fermentation condition including a pH within a range of 5.0 to 8.0, a temperature within a range of 25 to 50 ° C, and a stirring within a range of 50 to 500 rpm.

The culture medium used for the production of the culture concentrate of the Rizophus genus microorganism, the culture medium used for the production of the culture concentrate and the culture time and the culture time are the same and / or similar to those described above, and repeated explanation is avoided. And preferably Rhizopus oryzae .

The fermented vegetable oil according to the present invention is characterized in that the content of free essential unsaturated fatty acid is higher than that before fermentation by fermentation by the above method.

The free essential unsaturated fatty acids include gamma linolenic acid (GLA), linoleic acid (LA) and oleic acid (OA).

Gamma linolenic acid is a component of unsaturated fatty acids present only in women's breast milk and some plants, and is a necessary precursor to synthesize prostaglandins, known as active substances in the body. When the body lacks gamma linolenic acid, the physiologically active prostaglandin is not synthesized, so the necessary components for the normal functions of the body, including essential amino acids, are not well formed, resulting in an abnormality of the immune system. Therefore, the fermented oil of the present invention, which can solve the above problems, has an excellent effect of improving the itching and improving the atopic dermatitis as the content of the essential unsaturated fatty acid increases.

The above-mentioned free essential unsaturated fatty acid means a form in which the fatty acid is not bound to the glyceride. As the content of free essential unsaturated fatty acids increases, the stickiness decreases and the skin absorbency improves.

According to the present invention, there is provided a composition for external skin application for prevention of skin cell membrane damage, improvement of atopic dermatitis, improvement of itchiness of skin, moisturization, prevention and improvement of inflammatory skin trouble containing the above fermented vegetable oil as an effective ingredient; A cosmetic composition containing the above-mentioned fermented vegetable oil as an active ingredient for preventing skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement; And / or quasi-drugs for the prevention of damage to skin cell membranes, improvement of atopic dermatitis, improvement of skin itching, moisturizing, and prevention and improvement of inflammatory skin troubles, which contain the fermented vegetable oil as an active ingredient, may be provided. It is not intended to be intended to be used in the form of pharmaceutical preparations containing various kinds of pharmaceutical formulations containing the fermented vegetable oil having an increased content of essential unsaturated fatty acids as an effective ingredient and includes medicinal parts and quasi drugs which are applied to mammals, It will be understood by those skilled in the art that various types of products can be processed.

Further, in the method for producing a fermented vegetable oil according to the present invention, after the fermentation step, the fermented oil is centrifuged at 500 to 3000 rpm, and the concentrate of the microorganism of the present invention used for fermentation Can be separated by filtration to obtain a fermented vegetable oil which has been fermented. The obtained fermented vegetable oil may contain a large amount of essential unsaturated fatty acids.

The present invention also provides a pharmaceutical composition for external use for skin, which comprises the fermented vegetable oil according to the present invention as an active ingredient for prevention of skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement.

In addition, the present invention provides a cosmetic composition for prevention of skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement containing the fermented vegetable oil according to the present invention as an active ingredient.

The present invention also provides a quasi-drug for preventing damage to skin cell membranes, improving atopic dermatitis, improving skin itching, moisturizing, and preventing and treating inflammatory skin troubles, which comprises the fermented vegetable oil according to the present invention as an active ingredient.

The fermented vegetable oil according to the present invention is rich in free essential unsaturated fatty acids and has an inhibitory effect on NO production, COX-2 activity inhibition, inhibition of histamine production, inhibition of mast cell activation and degranulation inhibition, It can be effectively used as an effective ingredient of external preparation for skin, cosmetic composition or quasi-drug for improving itchiness and atopic dermatitis, moisturizing effect and regenerating damaged cell membrane.

The functional composition may be prepared in various formulations such as ointment for external use, cream, lotion, emulsion, lotion, essence, mist, gel, pack, oil, color cosmetics, soap, body wash, shampoo, rinse,

The functional composition may further include at least one or more additives selected from the group consisting of purified water, oil, a surfactant, a moisturizer, a stabilizer, an alcohol, a viscosity enhancer, a chelating agent, a coloring agent, an antiseptic,

Hereinafter, the present invention will be described in detail with reference to the following examples, but the scope of the present invention is not limited to the following examples.

Experimental Example 1 Comparative Experiment of Lipase Activity by Microorganism

Experiments were performed to compare the lipase activity of the culture concentrate of the Rizophus genus microorganism according to the present invention with other microorganisms.

As the control microorganisms, Burkholderia multivorans contains 1 wt% peptone, 0.5 wt% yeast extract, 0.02 wt% magnesium sulfate, 0.1 wt% potassium phosphate dibasic, 1 Pseudomonas fluorescence is a nutrient medium (nutrient medium) which is a type of bacterial medium which generally uses an extract of fish or meat meal, and mainly contains byproduct organic matter 0.5% by weight of peptone, 0.5% by weight of yeast extract, 0.8% by weight of salt, etc. may be added to 0.3% by weight of commercially available soup or especially for bacteriological extract to adjust the pH to about 7 1% olive oil was used for the culture medium, and Candida rugosa was used as a culture medium for YMB (1.5 g of yeast extract, 1.5 g of malt extract 1.5 g, peptone (2.5 g), dextrose (5.0 g), and distilled water (500 ml)) was added with olive oil to 1 wt% of the total amount of the medium. The medium of Pseudozima sp. Contains 0.01% glucose, 0.1% olive oil, 0.003% yeast extract, 0.003% malt extract, 0.003% peptone, 0.003% soybean flour, 0.02% ammonium sulfate , 0.001 wt% potassium phosphate, 0.005 wt% magnesium sulfate, 0.001 wt% calcium chloride, and 0.0001 wt% sodium chloride were used.

In order to measure the activity of lipase, a coloring method using a saponification reaction was used. The microorganisms were cultured, centrifuged at 12,000 rpm, and the supernatant was used as a crude enzyme solution. The substrate was incubated at 37 ° C for 30 minutes with 1% polyvinyl alcohol containing 10% olive oil, and the resulting free essential fatty acid was treated with copper-acetate- It reacts with pyridine (Cu-acetate-pyridine) to cause copper coloration. The degree of color development is measured at 715 nm. The lipase activity is expressed in units of units, and 1 U is defined as micromoles (μmol) of the fatty acid produced in one minute.

The highest pH and time of lipase activity according to each microorganism are shown in Table 1 below.

pH time Lipase activity
(U / ml) Rhizopus oryzae 6.0 120 4.3 Burkholderia multivorans (Burkholderia multivorans) 7.8 36 0.8 Pseudomonas fluorescence < RTI ID = 0.0 > 7.0 32 1.2 Candida rugosa 7.2 100 1.6 Pseudozima sp. 7.0 24 2.8

As shown in Table 1, it was confirmed that the Rhizopus orientalis used in the present invention exhibited the highest lipase activity by culturing.

Experimental Example 2 Comparative Experiment of Lipase Activity by Culture Time

The lipase activity was measured according to the incubation time of Rissofus oryzae exhibiting the highest lipase activity in Experimental Example 1 above. Experiments were conducted under the same culture conditions as in Experimental Example 1, except that the culture time was different. The results are shown in Table 2 below.

Strain Incubation time Lipase activity
(U / ml)
Rhizopus oryzae 90 hours 4.15 120 hours 4.30 140 hours 4.22

As shown in Table 2, in the case of Rizoffus oryzae, the lipase activity showed the highest activity at 120 hours of incubation, and then gradually decreased with time.

≪ Experimental Example 3 > Concentration of lipase produced from the culture solution and measurement of activity of lipase

The microorganisms were cultured, and centrifuged at 8000 rpm. Each of the cultures was treated with ammonium sulfate at a concentration of 20 to 100% by weight at 4 캜 to precipitate lipase. The supernatant was then removed by centrifugation at 4 ° C and 12,000 rpm for 20 minutes. The precipitate was dissolved in 0.05 M phosphate buffer (pH 7.0), and the concentrated lipase was obtained at 4 ° C using a dialysis membrane.

The activity of lipase according to the saturated concentration of ammonium sulfate used in the lipase precipitation is shown in the following table.

Ammonium sulfate concentration The lipase activity (U / ml) 0 to 20% 3.54 20 to 40% 10.22 40 to 60% 14.84 60 to 80% 15.33 80 to 100% 14.18

As can be seen from the above table, the activity of lipase was the highest when the saturation concentration of ammonium sulfate used in the lipase precipitation was 80 to 100%.

<Experimental Example 4> Measurement of yield of essential unsaturated fatty acid production according to fermentation temperature

In order to set the optimum temperature condition for the enzyme reaction, the temperature of the borage oil was set to 37 ° C, and the concentrated enzyme (using 60-80% ammonium sulfate) of Experimental Example 3 was added to phosphate buffer (Phosphate buffer). Then, the mixture was mixed at a ratio of 50% by weight of the enzyme solution to 50% by weight of borage oil and fermented at a rotation number of 300 rpm and a reaction temperature of 25 to 50 ° C for 72 hours.

Fermentation temperature
(° C) content (%) Gamma-linolenic acid
(γ-Linolenic acid) (GLA) Linoleic acid
Linoleic acid (LA) Oleic acid
(OA) 25 1.15 8.74 5.77 37 1.87 14.51 25.35 50 1.46 11.29 13.08

As a result of the experiment, the highest yield of essential unsaturated fatty acid was obtained at the fermentation temperature of 37 ° C.

<Experimental Example 5> Measurement of yield of essential unsaturated fatty acid production according to pH

In order to set the optimum pH condition for the enzyme reaction, the temperature of the borage oil was set to 37 ° C, and the concentrated enzyme (using 60-80% ammonium sulfate) of Experimental Example 3 was added to phosphate buffer (Phosphate buffer). Thereafter, the mixture was mixed at a ratio of 50% weight ratio of borage oil to 50% weight of enzyme solution, and fermented at a rotation number of 300 rpm and a reaction temperature of 37 캜 for 72 hours.

Fermentation pH content (%) Gamma-linolenic acid
(γ-Linolenic acid) (GLA) Linoleic acid
Linoleic acid (LA) Oleic acid
(OA) 5.0 0.94 8.12 9.74 6.0 1.33 10.66 13.55 7.0 1.87 14.51 25.35 8.0 1.42 10.61 17.35

The highest yield of essential unsaturated fatty acid was obtained at pH 7.0 of fermentation.

<Experimental Example 6> Measurement of yield of essential unsaturated fatty acid production by weight ratio of enzyme solution and oil

The concentration of enzyme (60-80% ammonium sulfate used) of Experimental Example 3 was adjusted to pH 5.0-7.0, after setting the temperature of the borage oil to 37 ° C in order to set the optimum weight ratio of the enzyme solution and the oil to which the enzyme reacted. 8.0 phosphate buffer (Phosphate buffer). Then, the enzyme solution was mixed at a weight ratio of 20% to 80% by weight of Borage Oil 80% to 20% by weight and fermented at a rotation number of 300 rpm and a reaction temperature of 37 ° C for 72 hours.

Enzyme solution: Oil
(Weight ratio,%) content (%) Gamma-linolenic acid
(γ-Linolenic acid) (GLA) Linoleic acid
Linoleic acid (LA) Oleic acid
(OA) 20: 80 1.35 11.51 17.68 50: 50 1.87 14.51 25.35 80: 20 1.08 6.53 10.35

Experimental results showed that the yield of essential unsaturated fatty acid production was the highest at 50: 50% by weight of enzyme solution and oil.

<Experimental Example 7> Measurement of yield of essential unsaturated fatty acid production according to fermentation time

To set the optimum fermentation time for the enzyme reaction, the temperature of the borage oil was set to 37 DEG C, and the concentrated enzyme (using 60-80% ammonium sulfate) of Experimental Example 3 was added to phosphate buffer solution (Sodium phosphate buffer) (50 mM). Thereafter, the mixture was mixed at a ratio of 50% weight ratio of Borax oil to 50% weight of enzyme solution, and fermented at a rotation number of 300 rpm and a reaction temperature of 37 캜 for 24 to 120 hours.

Fermentation time
(hour) content (%) Gamma-linolenic acid
(γ-Linolenic acid) (GLA) Linoleic acid
Linoleic acid (LA) Oleic acid
(OA) 24 0.64 3.54 8.52 48 1.26 9.52 16.27 72 1.87 14.51 25.35 96 1.96 14.85 26.72 120 2.05 15.03 27.38

The results showed that the longer the fermentation time, the higher the yield of essential unsaturated fatty acid production. However, the rate of production of essential unsaturated fatty acid decreased after 72 hours. Therefore, considering the economical aspect and efficiency of production of essential unsaturated fatty acid, the fermentation time is suitable for 72 hours.

PREPARATION EXAMPLE 1 Preparation of a Culture Concentrate of Risofussa Microorganism

Rhizopus oryzae as a microorganism of the genus Rhizopus was cultivated in PDB (Potato Dextrose Broth) containing 4% by weight of potato starch, 1% by weight of dextrose, 1% by weight of olive oil and water as a remainder, And 10% by volume relative to the culture medium, culturing under culture conditions of pH 6.0 and 30 ° C under aerobic conditions, followed by centrifugation at 1 to 10 ° C at 800 to 1200 rpm to obtain a precipitate obtained by salting out the supernatant obtained , Which was used as a culture concentrate of microorganisms of the genus Rizophus.

<Example 1 and Controls 1 and 2> Fermentation performance comparison between microorganisms (Pseudomonas sp. And Rythopus orientalis) and Rissofus orientalis culture concentrate

(Example 1) of the microorganism of the genus Rizopus according to the present invention of Preparation Example 1 (Example 1) and Pseudozima sp. Microorganism ( Pseudozima sp. ) (Control 1 ) And Rhizopus orientalis (control 2) were used to investigate the difference in fatty acid content when Borage oil was fermented as a vegetable oil. However, the culture concentrate of Rissofus oryzae was prepared by mixing the product obtained in Preparation Example 1 with a sodium phosphate buffer (50 mM) at pH 7.0.

The microorganisms Pseudojima and Rythopus orientus as fermented by the microorganisms themselves as control were inoculated into the PDB medium in an amount of 10 vol% relative to the medium volume, 50% by weight of Borage oil as a vegetable oil was mixed with 50% by weight of each of the Rissofus oryzae culture concentrates and fermented at 37 ° C for 72 hours at a rotation speed of 300 rpm. The fermented oil was fermented at 4000 rpm for 20 minutes After centrifugation, it was filtered to obtain a fermented vegetable oil. The fatty acid content after fermentation was measured, and the results are shown in Table 8 below.

content (%) Gamma-linolenic acid
(γ-Linolenic acid) (GLA) Linoleic acid
Linoleic acid (LA) Oleic acid
(OA) Control 1 0.95 7.19 12.82 Control 2 0.58 4.29 7.82 Example 1 1.87 14.51 25.35

As shown in Table 8, when fermented with a culture concentrate of Rissofus oryzae according to the present invention, fermentation of the same Borage oil resulted in higher fatty acid content than fermentation with the microorganism itself About twice as much) as the fermented vegetable oil.

In addition, the fermented vegetable oils obtained in Example 1 and Controls 1 and 2 were subjected to sensory evaluation. The sensory evaluation of odor was carried out on 15 adult males and females, and the fermented vegetable oils of Example 1 and Controls 1 and 2 were divided into three times. The evaluation vessels were the same and were evaluated by providing 50 ml each. The samples were marked so that they could not be seen by the evaluators, and then the degree of smell was evaluated. The scores were rated 1 to 6, very poor to 1, and very good to 6, and the results are averaged and shown in Table 9 below.

Rating Control 1 2.65 ± 0.6 Control 2 2.5 ± 1.5 Example 1 4.4 ± 0.9

As shown in Table 9, when the fermented product was fermented with the culture concentrate according to the present invention, the resistance to odor was less than that of the control 2 fermented directly with microorganisms.

&Lt; Examples 2 to 4 >

(Example 2), evening primrose oil (Example 3) and camellia oil (Example 4) as the vegetable oil to be fermented according to the present invention were used in the same manner as in Example 1, And fermented with a culture concentrate.

The essential fatty acid content of each of the fermented vegetable oils obtained in Examples 1 to 4 was measured, and the results are shown in Table 10 below.

Oil type Content (mg / g) Gamma-linolenic acid
(γ-Linolenic acid) (GLA) Linoleic acid
Linoleic acid (LA) Oleic acid
(OA) Example 1 1.87 14.51 25.35 Example 2 1.35 12.86 6.82 Example 3 1.21 15.13 32.65 Example 4 2.02 20.44 9.07

&Lt; Comparative Examples 3 to 6 >

(Comparative Example 3), unfermented moringa oil (Comparative Example 4), unfermented evening primrose seed oil (Comparative Example 5), and unfermented camellia oil (Comparative Example 3) 6) were used.

<Experimental Example 8> Confirmation effect of inhibiting the production of skin inflammatory substance (NO)

In order to confirm the effect of inhibiting the production of nitric oxide (NO), which is a skin inflammation inducing substance, in samples of the respective Examples, nitrite (nitrite) in the culture liquid produced by treating LPS (lipopolysaccharide) nitrite concentrations were measured using a Griess reaction.

Specifically, a 96-well culture plate was coated with 1 × 10 ⁴ cells / well in a 96-well culture plate using DMEM medium (Dulbecco's Modified Eagle's Medium: Dulbecco's modified Eagle medium) containing FBS and antibiotics RAW 264.7 cells derived from macrophages were cultured at 37 ° C. in a 5% CO 2 incubator for 24 hours after dispensing. After 24 hours, the medium used for the previous culture was removed and a new DMEM medium without FBS and antibiotics was dispensed and each sample was treated. After 1 hour, 1 占 퐂 / ml of LPS was treated and cultured for 24 hours. The NO generated during the culture was measured using the Griess reagent and the total NO2- concentration in the cell culture. The cell culture supernatant (50 μl) and the Griess reagent (50 μl) were mixed and reacted in 96-well plates for 10 min. Absorbance was measured at 540 nm using an ELISA reader (enzyme immunoassay reader).

The results are shown in FIG. 1 and Table 11.

NO (nitric oxide) production (μM) sham 37.66 + - 7.16 (-) control group 387.94 + 56.83 Comparative Example 1 280.11 + - 16.44 Comparative Example 2 318.73 ± 36.51 Comparative Example 3 364.25 + - 4.82 Comparative Example 4 339.24 + 30.94 Example 1 64.52 + - 6.02 Example 2 88.91 + - 16.42 Example 3 105.94 ± 26.88 Example 4 124.06 ± 8.83

As a result of the experiment, it was found that the use of the oils prepared according to Examples 1, 2, and 3 to 4 of the present invention markedly inhibited the production of NO, which is one of the skin inflammation inducing substances induced by LPS in mouse macrophages ( 1 and Table 11).

<Experimental Example 9> Determination of the inhibitory effect of COX-2

To investigate the effect of each sample on the activity of COX-2 (cyclooxygenase-2), a skin inflammation inducing enzyme, arachidonic acid, which is a substrate of this enzyme, was used to induce oxidation of hematin And the discoloration was measured using the phenomenon.

Specifically, 10 μl of COX-2 enzyme (30 units / mL), heme solution (150 μM / mL), and 10 μl of COX-2 enzyme were added to a 1.5 mL tube to cause the enzymatic reaction of COX-2 producing prostaglandin using arachidonic acid as a substrate , 950 μl of reaction buffer (0.1 M Tris-HCl, 5 mM EDTA, 2 mM phenol, pH 8.0) was added, and 20 μl of the sample diluted by sequential concentration was added thereto, followed by stabilization at 37 ° C. for 5 minutes. 10 μl of arachidonic acid was added thereto, followed by reaction at 37 ° C for 2 minutes, and then 50 μl of 1M HCl was added to stop the reaction. 100 μl of SnCl 2 solution (50 mg / ml) was added and mixed well, followed by reaction at room temperature for 5 minutes.

In order to perform competitive ELISA with the prostaglandin produced in the above reaction, the reaction solution was diluted 2000-fold and 4000-fold with EIA buffer, and 50 μl was added to 96-well plates coated with prostaglandin antibody. Then, 50 μl of PGs screening AChE tracer 50 μl of antiserum was added and reacted at room temperature for 18 hours. The 96 wells were washed 5 times with buffer, 200 ㎕ of Ellman's reagent was added, and developed for 60 minutes and then absorbance was measured at a wavelength of 410 nm.

The degree of inhibition of COX-2 activity was expressed by the value of IC50 (ppm), which is a 50% inhibitory concentration according to the manufacturer's method. Experimental results were expressed as mean ± standard deviation by repeating twice.

The graph shows the IC50 value in ppm of the oil produced by Examples 1, 2, 3 to 4 and Comparative Examples 1, 2 and 3 to 4 of the present invention inhibiting COX-2 enzyme by 50%.

The results are shown in FIG. 2 and Table 12.

IC ₅₀ of COX-2 (ppm) Comparative Example 1 316.49 + - 53.64 Comparative Example 2 264.22 + - 39.49 Comparative Example 3 294.16 + - 8.66 Comparative Example 4 285.42 + - 29.48 Example 1 164.94 ± 11.38 Example 2 155.82 + - 5.51 Example 3 167.51 ± 27.94 Example 4 159.03 + - 14.67

As a result, it was found that the activity of COX-2, a skin inflammation inducing enzyme, was significantly inhibited when the oils prepared in Examples 1, 2, and 3 to 4 of the present invention were used (FIG. 2 and Table 12).

<Experimental Example 10> Confirmation of inhibitory effect on histamine formation

In order to measure the inhibition of the production of histamine, which is a skin irritant and skin inflammation inducer, mast cells isolated from abdominal cavity of SD rats were treated with compound 48/80, a degreasing inducer, The amount of histamine was measured by ELISA.

Specifically, the SD rats were sacrificed by ether anesthesia, and then incubated with HEPES-Tyrode buffer (136 mM NaCl, 5 mM KCl, 2 mM CaCl2, 0.6 mM NaH2PO4, 2.75 mM MgCl2, 5.4 mM HEPES, , 1.0 mg / mL Glucose, pH 7.4) was injected into the abdominal cavity of the rats, and the abdominal wall was lightly massaged for 90 seconds. The abdominal wall was cut and the peritoneal lavage fluid was collected with a syringe and centrifuged at 200 × g for 10 minutes at 4 ° C. The supernatant was discarded and resuspended to a density of 1 × 10 6 cells / mL in HEPES-Tyrode buffer. Add 0.75 mL of cell suspension and 0.5 mL of HEPES-Tyrode buffer to 3.5 mL of Isotonic Percol solution (10 mL of Hank's solution + 9 mL of Percoll), centrifuge at 125 x g for 15 minutes at 4 ° C, discard the supernatant, buffer to make a pure mast cell suspension. Separated mast cells were suspended in HEPES-Tyrode buffer to a concentration of 5 × 10 4 cells / mL, and each sample was treated. After 10 minutes, compound 48/80 (1 μg / mL) was treated with positive control (compound 48/80) and test group (negative) except for negative control (PBS treated group) and reacted for 20 minutes. The supernatant was obtained by centrifugation at 400 xg for 10 min for histamine release from cells. To determine the rate of inhibition of histamine release by supernatant, the histamine assay kit was used to determine the competitive ELISA method as follows. Standard histamine and samples of each concentration were dispensed in 50 μL into a 96-well plate coated with antihistamine antibody, and 50 μL of an enzyme conjugate was added, followed by reaction at room temperature for 45 minutes. After washing four times with buffer, 150 μl of enzyme substrate was added, reacted at room temperature for 30 minutes, and absorbance was measured at 650 nm using an ELISA plate reader. The amount of histamine secreted (ng / mL) was calculated by substituting the ratio of the liberated histamine to the antihistamine antibody (absorbance of experimental group / absorbance of maximum enzyme conjugate group X 100) into histamine standard curve provided by the ELISA kit manufacturer .

The results are shown in FIG. 3 and Table 13.

Histamine secreted from the medium (ng / ml) sham 14.64 ± 2.49 (-) control group 51.85 ± 6.16 Comparative Example 1 39.15 + - 2.42 Comparative Example 2 44.16 + - 4.55 Comparative Example 3 42.64 ± 1.05 Comparative Example 4 41.55 ± 3.88 Example 1 28.45 + - 1.15 Example 2 32.81 ± 1.52 Example 3 35.11 ± 5.43 Example 4 33.47 ± 1.68

Experimental results showed that the use of the oils prepared in Examples 1, 2, and 3 to 4 of the present invention significantly inhibited the skin itching and the activity of histamine, which is a skin inflammation-inducing substance (FIG. 3 and Table 13).

<Experimental Example 11> Confirmation of degranulation inhibition of mast cells

IgE binds to FcεRI, a receptor for IgE present on the surface of mast cells, leading to the degranulation of mast cells and releasing histamine stored in intracellular granules. This liberated histamine causes vasodilatation and muscle contraction resulting in immediate allergic symptoms.

Mast cells isolated from the abdominal cavity of SD rats were used in the same manner as in the measurement of inhibition of histamine release of mast cells. Separated mast cells were suspended in HEPES-Tyrode buffer to a concentration of 5 × 10 4 cells / mL, and each sample was treated. Ten minutes later, the positive control group and the sample treatment group except for the negative control (PBS treatment) were treated with compound 48/80 (1 ㎍ / mL) and reacted for 20 minutes. 20 μl of abdominal suspension containing the reacted mast cells is placed on a slide glass and allowed to stand at room temperature for 10 minutes so that mast cells can be precipitated. Mast cells were observed under an optical microscope at a magnification of 400 times and classified into a normal type and a degranulation type.

The results are shown in Figs. 4, 5, and 14.

Histamine secreted from the medium (ng / ml) sham 5.16 ± 0.54 (-) control group 78.14 + - 7.52 Comparative Example 1 72.84 + 9.33 Comparative Example 2 69.62 + - 11.52 Comparative Example 3 70.06 + - 2.85 Comparative Example 4 75.45 + - 8.85 Example 1 25.11 + - 3.54 Example 2 22.97 + - 6.99 Example 3 18.49 + - 2.57 Example 4 35.86 ± 1.05

As a result of the experiment, it was found that when oil prepared in Examples 1, 2, and 3 to 4 of the present invention was used, the activation and degranulation of mast cells inducing the release of histamine, which is a skin itching and skin inflammation inducing substance, was remarkably suppressed (Figs. 4, 5 and 14).

<Experimental Example 12> Cell (3T3) safety measurement

EXAMPLES OF THE INVENTION MTT assay was performed to measure the toxicity of 3T3 cells, the mouse fibroblast cells of the oil prepared according to 1, 2, and 3 to 4.

Specifically, toxicity to mouse fibroblasts was assayed by measuring MTT [(3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (Sigma, MO, USA)]. 3T3 cells were plated at 1 × 104 cells / mL per well in 96 wells and cultured at 37 ° C and 5% CO2 for 24 hours. After 24 hours, the medium used for the previous culture was removed, the sample was dissolved in DMSO, and diluted with the medium so that the final concentration of DMSO was 1%. The concentration of the extract was 5 μg / Hour and 24 hours, respectively. After the treatment, 20 μl of 0.2% MTT solution was added to each well and reacted for 3 hours at 37 ° C in a 5% CO2 incubator. After the reaction, the supernatant was removed, and 150 μl of DMSO was added. All of the formazan produced at room temperature for 10 minutes was dissolved, and the absorbance at 570 nm was measured using an ELISA reader. To investigate the effects on DMSO, experiments were performed by dividing into a control group and a control group for 1% DMSO. Cell viability (%) was expressed by the following equation.

Cell survival rate (%) = (absorbance of sample treated group / absorbance of control group) x 100

The results are shown in Figs. 6 to 9.

As a result, it was found that the cytotoxicity of the oil prepared in Examples 1, 2, and 3 to 4 of the present invention was not toxic at a concentration of 500 ppm or less (FIGS. 6 to 9).

<Experimental Example 13> Measurement of skin irritation

EXAMPLES In order to measure the degree of skin irritation of the oil prepared according to Examples 1, 2, and 3 to 4, a sample was applied to an area of 3 × 3 ㎠ in diameter of the inside of the arm, And 48 hours later.

Evaluation of skin irritability The method of calculating the numerical value and the measurement results are shown in Tables 15 and 16 below.

Degree of stimulation Rating No stimulation 10 Very weak stimulus 8 Weak stimulation 6 Normal stimulus 4 Slightly strong stimulus 2 Strong stimulus 0

division 24 hours 48 hours Control group 9.8 ± 0.6 9.7 ± 1.2 Example 1 9.6 ± 1.6 9.8 ± 1.4 Example 2 9.9 ± 2.5 10.0 0.0 Example 3 10.0 0.0 9.8 ± 1.4 Example 4 9.8 ± 1.2 9.6 ± 0.6

As a result of the experiment, it can be seen from the table that there is no skin irritation to the oil prepared by Examples 1, 2, 3 to 4 prepared according to the present invention (Table 15 and Table 16).

<Experimental Example 14> Measurement of change in fatty acid content over time during direct fermentation of microorganisms

As a reference experiment for the present invention, a microorganism culture solution (Ricofus oryzae not concentrated, which was not concentrated) was cultured in the same manner as in Synthesis Example 1 except that the concentrate was not concentrated At this time, the fermentation time was further increased as compared with the present invention, and the change in fatty acid content was measured. The results are shown in FIGS. 17 and 10.

Fermentation time using microbial culture content (%) Gamma-linolenic acid
(γ-Linolenic acid) (GLA) Linoleic acid
Linoleic acid (LA) Oleic acid
(OA) 24 0.16 1.46 2.25 48 0.25 2.15 4.57 72 0.58 4.29 7.82 96 0.62 4.35 7.92 120 0.71 4.75 8.01

As shown in Table 17 and FIG. 10, it was confirmed that when the non-concentrated microorganism culture solution was used, no significant change in the fatty acid content was observed after about 72 hours.

Claims (15)

Which is obtained by fermenting a vegetable oil selected from the group consisting of Borage oil, Moringa oil, Evening Primrose oil, Camellia oil, Olive oil, Sunflower seed oil and a mixture of two or more thereof with a culture concentrate of Rizophus genus microorganism A characteristic fermented vegetable oil. The method according to claim 1,
The culture concentrate of the Rizophus genus microorganism is inoculated with the microorganism of the genus Rizophus in an amount of 10 vol.% Relative to the medium volume and cultured under culture conditions at pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, Wherein the fermented vegetable oil is a precipitate obtained by salting out a supernatant obtained by centrifuging at 1 to 10 DEG C at 800 to 1200 rpm. 3. The method of claim 2,
Wherein the culture medium is PDB (Potato Dextrose Broth) comprising 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose and water as a balance. The method of claim 3,
Wherein the PDB further comprises 0.1 to 10% by weight of oil based on the total weight of the PDB. 3. The method of claim 2,
Wherein the fermented vegetable oil has a culture time of 80 to 150 hours under the culture conditions. The method according to claim 1,
Wherein the fermentation of the vegetable oil is carried out by adding a culture concentrate of the microorganism of the genus Rizophus as a remainder to 10 to 80% by weight of a vegetable oil preheated to a temperature within a range of 10 to 40 DEG C, At a temperature within a range of 25 to 50 DEG C, and within a range of 50 to 500 rpm. The method according to claim 1,
Wherein the microorganism in Rizophus is Rhizopus oryzae. A method for producing a fermented vegetable oil by fermenting a vegetable oil,
(1) a step of adding a culture concentrate of the microorganism of the genus Rizophus as a remainder to 10 to 80% by weight of a vegetable oil preheated to a temperature within a range of 10 to 40 占 폚; And
(2) a culture step of culturing at a pH within a range of 5.0 to 8.0, a temperature within a range of 25 to 50 占 폚, and a culture condition containing agitation within a range of 50 to 500 rpm;
/ RTI &gt;
Wherein the vegetable oil is selected from the group consisting of borage oil, moringa oil, evening primrose oil, camellia oil, olive oil, sunflower seed oil and mixtures of two or more thereof. 9. The method of claim 8,
The culture concentrate of the Rizophus genus microorganism is inoculated with the microorganism of the genus Rizophus in an amount of 10 vol.% Relative to the medium volume and cultured under culture conditions at pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, Wherein the supernatant is obtained by salting out the supernatant obtained by centrifugation at 800 to 1200 rpm at 1 to 10 &lt; 0 &gt; C. 10. The method of claim 9,
Wherein the culture medium is PDB (Potato Dextrose Broth) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose, and water as a remaining amount. 10. The method of claim 9,
Wherein the culturing time is in the range of 80 to 150 hours under the culturing conditions. 9. The method of claim 8,
Wherein the microorganism in Rizophus is Rhizopus oryzae. A pharmaceutical composition for dermatological use for prevention of damage to skin cell membrane, improvement of atopic dermatitis, improvement of itchiness of skin, moisturization, prevention and improvement of inflammatory skin trouble containing fermented vegetable oil according to any one of claims 1 to 7 as an effective ingredient . A cosmetic composition for preventing skin cell membrane damage, atopic dermatitis improvement, skin itching improvement, moisturizing, inflammatory skin trouble prevention and improvement containing the fermented vegetable oil according to any one of claims 1 to 7 as an active ingredient. A quasi-drug for preventing damage to skin cell membranes, improving atopic dermatitis, improving skin itching, moisturizing, and preventing and treating inflammatory skin troubles, which comprises the fermented vegetable oil according to any one of claims 1 to 7 as an active ingredient.

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